• Asian-Australasian Journal of Animal Sciences
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• v.21 no.3
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• pp.392-403
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• 2008

Four ruminally fistulated Hanwoo steers were used to determine the effects of level and degradability of dietary protein on ruminal fermentation, blood metabolites and concentration of soluble non-ammonia nitrogen (SNAN) in ruminal (RD) and omasal digesta (OD). Experiments were conducted in a $4{\times}4$ Latin square design with a $2{\times}2$ factorial arrangement of treatments. Factors were protein supplements with two ruminal crude protein (CP) degradabilities, corn gluten meal (CGM) that was low in degradability (rumen-degraded protein (RDP), 23.4% CP) or soybean meal (SBM) that was high in degradability (RDP, 62.1% CP), and two feeding levels of CP (12.2 or 15.9% dry matter). Ruminal fermentation rates and plasma metabolite concentrations were determined from the RD collected at 2-h intervals and from the blood taken by jugular puncture, respectively. The SNAN fractions (free amino acid, peptide and soluble protein) in RD and OD collected at 2-h intervals were assessed by ninhydrin assay. Mean ruminal ammonia concentrations were 40.5, 74.8, 103.4 and 127.0 mg/L for low CGM, high CGM, low SBM and high SBM, respectively, with statistically significant differences (p<0.01 for CP level and p<0.001 for CP degradability). Blood urea nitrogen concentrations were increased by high CP level (p<0.001) but unaffected by CP degradability. There was a significant (p<0.05) interaction between level and degradability of CP on blood albumin concentrations. Albumin was decreased to a greater extent by increasing degradability of low CP diets (0.26 g/dl) compared with high CP diets (0.02 g/dl). Concentrations of each SNAN fraction in RD (p<0.01) and OD (p<0.05) for high CP diets were higher than those for low CP diets, except for peptides but concentrations of the sum of peptide and free amino acid in RD and OD were significantly higher (p<0.05) for high CP diets than for low CP diets. Soybean meal diets increased free amino acid and peptide concentrations in both RD (p<0.01) and OD (p<0.05) compared to CGM diets. High level and greater degradability of CP increased (p<0.001) mean concentrations of total SNAN in RD and OD. These results suggest that RDP contents, increased by higher level and degradability of dietary protein, may increase release of free amino acids, peptides and soluble proteins in the rumen and omasum from ruminal degradation and solubilization of dietary proteins. Because SNAN in OD indicates the terminal product of ruminal metabolism, increasing CP level and degradability appears to increase the amount of intestine-available nitrogen in the liquid phase.

• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.186-195
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• 2015

Anthranilate synthase (AS) is a key enzyme in the biosynthesis of tryptophan (Trp), which is the precursor of bioactive metabolites like indole-3-acetic acid and other indole alkaloids. Alpha anthranilate synthase 2 (OsASA2) plays a critical role in the feedback inhibition of tryptophan biosynthesis. In this study, two vectors with single (F124V) and double (S126F/L530D) point mutations of the OsASA2 gene for feedback-insensitive ${\alpha}$ subunit of rice anthranilate synthase were constructed and transformed into wildtype Dongjinbyeo by Agrobacterium-mediated transformation. Transgenic single and double mutant lines were selected as a single copy using TaqMan PCR utilized nos gene probe. To select intergenic lines, the flanking sequence of RB or LB was digested with a BfaI enzyme. Four intergenic lines were selected using a flanking sequence tagged (FST) analysis. Expression in rice (Oryza sativa L.) of the transgenes resulted in the accumulation of tryptophan (Trp), indole-3-acetonitrile (IAN), and indole-3-acetic acid (IAA) in leaves and tryptophan content as a free amino acid in seeds also increased up to 30 times relative to the wildtype. Two homozygous event lines, S-TG1 and D-TG1, were selected for characterization of agronomic traits and metabolite profiling of seeds. Differentially expressed genes (DEGs), related to ion transfer and nutrient supply, were upregulated and DEGs related to co-enzymes that work as functional genes were down regulated. These results suggest that two homozygous event lines may prove effective for the breeding of crops with an increased level of free tryptophan content.

• Archives of Pharmacal Research
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• v.28 no.7
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• pp.816-822
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• 2005

Catharsius protease-2 (CPM-2) was isolated from the body of dung beetles, Catharsius molossus, using a three step purification process (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and affinity chromatography on DEAE Affi-Gel blue). The purified CPM-2, having a molecular weight of 24 kDa, was assessed homogeneously by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of CPM-2 was composed of X Val Gin Asp Phe Val Glu Glu lie Leu. CPM-2 was inactivated by $Cu^{2+}\;and\;Zn^{2+}$ and strongly inhibited by typical serine proteinase inhibitors such as TLCK, soybean trypsin inhibitor, aprotinin, benzamidine, and ${\alpha}_1$-antitrypsin. However, EDTA, EGTA, cysteine, $\beta$-mercaptoethanol, E64, and elastatinal had little effect on enzyme activity. In addition, antiplasmin and antithrombin III were not sensitive to CPM-2. Based on the results of a fibrinolytic activity test, CPM-2 readily cleaved $A{\alpha}-$ and $B{\beta}$-chains of fibrinogen and fibrin, and y-chain of fibrinogen more slowly. The nonspecific action of the enzyme resulted in extensive hydrolysis, releasing a variety of fibrinopeptides of fibrinogen and fibrin. Polyclonal antibodies of CPM-2 were reactive to the native form of antigen. The ELISA was applied to detect quantities, in nanograms, of the antigen in CPM-2 protein.

• Journal of Crop Science and Biotechnology
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• v.10 no.2
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• pp.112-116
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• 2007

Kinetin(KN) is an inducer of rice(Oryza sativa L.) defense/stress responses, as evidenced by the induction of inducible secondary metabolite and defense/stress protein markers in leaf. We show a novel light-dependent effect of KN-triggered defense stress responses in rice leaf. Leaf segments treated with KN(100 ${\mu}M$) show hypersensitive-like necrotic lesion formation only under continuous light illumination. Potent accumulation of two phytoalexins, sakuranetin and momilactone A(MoA) by KN that peaks at 48 h after treatment under continuous light is completely suppressed by incubation under continuous dark. Using two-dimensional gel electrophoresis we identified KN-induced changes in ribulose-1, 5-bisphosphate carboxylase/oxygenase, energy- and pathogenesis-related proteins(OsPR class 5 and 10 members) by N-terminal amino acid sequencing and mass spectrometry. These changes were light-inducible and could not be observed in the dark(and control). Present results provide a new dimension(light modulation/regulation) to our finding that KN has a potential role in the rice plant self-defense mechanism.

• Journal of Ginseng Research
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• v.45 no.6
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• pp.726-733
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• 2021

Background: Panax notoginseng is a highly valued medicinal herb used widely in China and many Asian countries. Its root and rhizome have long been used for the treatment of cardiovascular and hematological diseases. Imaging the spatial distributions and dynamics of metabolites in heterogeneous plant tissues is significant for characterizing the metabolic networks of Panax notoginseng, and this will also provide a highly informative approach to understand the complex molecular changes in the processing of Panax notoginseng. Methods: Here, a high-sensitive MALDI-MS imaging method was developed and adopted to visualize the spatial distributions and spatiotemporal changes of metabolites in different botanical parts of Panax notoginseng. Results: A wide spectrum of metabolites including notoginsenosides, ginsenosides, amino acids, dencichine, gluconic acid, and low-molecular-weight organic acids were imaged in Panax notoginseng rhizome and root tissues for the first time. Moreover, the spatiotemporal alterations of metabolites during the steaming of Panax notoginseng root were also characterized in this study. And, a series of metabolites such as dencichine, arginine and glutamine that changed with the steaming of Panax notoginseng were successfully screened out and imaged. Conclusion: These spatially-resolved metabolite data not only enhance our understanding of the Panax notoginseng metabolic networks, but also provide direct evidence that a serious of metabolic alterations occurred during the steaming of Panax notoginseng.

• Journal of Microbiology and Biotechnology
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• v.29 no.8
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• pp.1212-1220
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• 2019

The study of metabolomics in natural products using the diverse analytical instruments including GC-MS, LC-MS, and NMR is useful for the exploration of physiological and biological effects and the investigation of drug discovery and health functional foods. Cordyceps militaris has been very attractive to natural medicine as a traditional Chinese medicine, due to its various bioactive properties including anti-cancer and anti-oxidant effects. In this study, we analyzed the metabolite profile in 50% ethanol extracts of C. militaris fruit bodies from three development periods (growth period, matured period, and aging period) using $^1H-NMR$, and identified 44 metabolites, which are classified as 16 amino acids, 10 organic acids, 5 carbohydrates, 3 nucleotide derivatives, and 10 other compounds. Among the three development periods of the C. militaris fruit body, the aging period showed significantly higher levels of metabolites including cordycepin, mannitol (cordycepic acid), and ${\beta}-glucan$. Interestingly, these bioactive metabolites are positively correlated with antitumor growth effect; the extract of the aging period showed significant inhibition of HepG2 hepatic cancer cell proliferation. These results showed that the aging period during the development of C. militaris fruit bodies was more highly enriched with bioactive metabolites that are associated with cancer cell growth inhibition.

• Journal of Microbiology and Biotechnology
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• v.27 no.5
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• pp.925-932
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• 2017

Changes in the metabolite profiles of Lactobacillus sakei and its growth media, based on different culture times (0, 6, 12, and 24 h), were investigated using gas chromatography-mass spectrometry (MS) and liquid chromatography-MS with partial least squares discriminant analysis, in order to understand the growth characteristics of this organism. Cell and media samples of L. sakei were significantly separated on PLS-DA score plots. Cell and media metabolites, including sugars, amino acids, and organic acids, were identified as major metabolites contributing to the difference among samples. The alteration of cell and media metabolites during cell growth was strongly associated with energy production. Glucose, fructose, carnitine, tryptophan, and malic acid in the growth media were used as primary energy sources during the initial growth stage, but after the exhaustion of these energy sources, L. sakei could utilize other sources such as trehalose, citric acid, and lysine in the cell. The change in the levels of these energy sources was inversely similar to the energy production, especially ATP. Based on these identified metabolites, the metabolomic pathway associated with energy production through lactic acid fermentation was proposed. Although further studies are required, these results suggest that MS-based metabolomic analysis might be a useful tool for understanding the growth characteristics of L. sakei, the most important bacterium associated with meat and vegetable fermentation, during growth.

• Korean Journal of Food Science and Technology
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• v.51 no.3
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• pp.248-257
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• 2019

This study aimed to determine the chemical components of different cultivars (Gabjubaekmok, Sangjudungsi, and Godongsi) of persimmon (Diospyros kaki Thunb.) and the effects of maturity. The detected free sugars were fructose and glucose, and glucose levels tended to increase with maturity. Palmitic acid, oleic acid, linoleic acid and stearic acid, and potassium were major components. In the amino acid component analysis, the following were usually present in the following month: glutamic acid in Gabjubaekmok in July, lysine in Sangjudungsi in August, and threonine in Godongsi in October. Vitamin C tended to increase with aging, and fruit harvested in July had the highest total phenolic and flavonoid contents. In the metabolite analysis, there were significant differences among cultivars and with maturity. The major physiological compounds were analyzed using ultra performance liquid chromatography quadrupole time-of-flight tandem mass ($UPLC/Q-TOF/MS^2$) and were citric acid and gallic aicd. As maturity progressed, citric acid increased but gallic acid decreased.

• Journal of Bio-Environment Control
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• v.31 no.3
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• pp.170-179
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• 2022

Glutamic acid is a precursor of essential amino acids that play an important role in plant growth and development. It is one of the biostimulants that reduce cold stress damage by stimulating biosynthetic pathways leading to cryoprotectants. This study evaluated the effects of glutamic acid foliar application on Kimchi cabbage under low-temperature stress. There were six treatments, combining three photo-/dark periods temperature levels (11/-1℃ extremely low, E; 16/4℃ moderately low, M; and 21/9℃ optimal, O) with and without glutamic acid foliar application (0 and 10 mg·L^-1; Glu 0 and Glu 10). Glutamic acid foliar application was sprayed once 10 days after transplanting, and then temperature treatment immediately after glutamic acid foliar application was conducted for up to four days. After four days of treatment, abscisic acid (ABA), phaseic acid (PA), dihydrophaseic acid (DPA), and abscisic acid-glucose ester (ABA-GE) contents were higher with Glu 10 treatment than Glu 0 treatment in M treatment. Glucose content was highest in E with Glu 10 treatment (52.1 mg·100 g^-1 dry weight), while fructose content was highest in O with Glu 0 treatment (134.6 mg·100 g^-1 dry weight). The contents of glucolepiddin (GLP), glucobrassicin (GBS), 4-methoxyglucobrassicin (4MGBS), neoglucobrassicin (GNBS), and gluconasturtiin (GNS) were highest among all treatments in E with Glu 10 treatments (0.72, 2.05, 1.67, 9.40 and 0.85 µmol·g^-1 dry weight). After two days of treatment, rapid changes in PA and DPA contents of E with Glu 10 treatments were confirmed, and several individual glucosinolate contents (GLP, GBS, 4MGBS, GNBS, and GNS) were significantly different depending on low temperature and glutamic acid treatment. In addition, the content of fructose was significantly lower than that of O treatment in E and M treatments after four days of treatment. Therefore, although the changes in PA, DPA, glucose, fructose, and individual glucosinolates according to low temperature and glutamic acid foliar treatment were shown. A clear correlation between low temperature and glutamic acid effects could not be evaluated. Results indicated that Brassica crops are cryophilic vegetables, do not react sensitively to low temperatures, and mostly have cold resistance.

• The Korean Journal of Pesticide Science
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• v.10 no.2
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• pp.76-83
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• 2006

In order to investigate the impacts of non-selective herbicide, glufosinate-ammonium (ammonium 4-[hydroxy(methyl)phosphinoyl] -DL-homoalaninate, GLA) to the non-target organisms, earthworm was exposed to GLA in the field soil for a month, and microbial populations in the soil were investigated after application of GLA. Simultaneously, the residues of GLA and its metabolite, 3-MPP were analyzed in the same soil. Meanwhile, to elucidate the influence of GLA to the growth of non-target crops incase of inter-furrow application, the amounts of carotenoid, chlorophyll, amino acid, proteins and sugars in the leaves of potato and chinese cabbage grown in the same field were investigated. In result, the dead earthworm was not observed during the test period, and the increasing rates of bodyweight were $9.410{\sim}11.603%$ in GLA-treated plots and 5.645% in GLA-untreated plots. The populations of fungi, bacteria and actinomycetes in the GLA-treated soils were $6.2{\times}10^4$, $1.5{\times}10^6$ and $5.7{\times}10^4$, respectively. They maintained relatively similar levels to the control which were $3.7{\times}10^4$, $3.7{\times}10^5$ and $3.7{\times}10^4$, respectively. In residue analysis, the limit of detection of GLA was 0.02 mg $kg^{-1}$, that of 3-MPP was the same level, and the half-life of GLA was 15 days in sandy clay loam soil. This result indicates that GLA was degraded very quickly in field soil. On the other hand, the amounts of physiological, biochemical components such as carotenoid, amino acid, chlorophyll, protein and sugar were ranged from 90.0 to 104.3% in potato and from 99.0 to 112.7% in chinese cabbage. Comparing with hand-weeded plots, it is indicated that GLA had not affected to the growth of non-target crops when applied at inter-furrow in crops-growing field.